The reliable determination of the Ag(I) affinity for biomolecules is an essential issue in the fields of structural analysis and sensor design. However, the urgent problem confronting researchers is lack of a direct and accurate Ag(I) affinity evaluation as a reference standard for ligand analysis. We communicated here a straightforward and high-efficiency method of measuring Ag(I) affinity exactly on the basis of the unique calculation algorithm and the design of a special peptide RFPRDD (P) as Ag(I) binding motif. According to UV–vis competition between the corresponding complexes (AgP) and biomolecules (peptides, amino acids and ssDNA), the decrease of the signature at 300 nm characteristic of AgP was obtained for quantitative analysis. The primary advantages of this strategy were the widespread application, high accuracy and reference significance, which were corroborated by theoretical calculations. To identify its potential in biosensing, two kinds of testing models for Ag(I) were proposed by AgBP2-decorated and Ag4-decorated gold nanoparticles, the detection limits of which were 2 nM and 75 nM respectively. By contrast of the sensing property of the functional peptides (AgBP2, Ag4), we afforded evidence that this conception could be regarded as an evaluation criterion for the selection and performance optimization of sensitive elements, thereby holding a dominant position in the biosensors.

Ag（I）对生物分子亲和力的可靠测定是结构分析和传感器设计领域的重要问题。然而，研究人员面临的紧迫问题是缺乏直接和准确的Ag（I）亲和力评估作为配体分析的参考标准。我们在此交流了一种直接且高效的方法，正是基于独特的计算算法以及作为Ag（I）结合基序的特殊肽RFPRDD（P）的设计，准确地测量了Ag（I）亲和力。根据相应复合物（AgP）与生物分子（肽，氨基酸和ssDNA）之间的紫外可见竞争，定量分析发现AgP在300 nm处的特征减少。该策略的主要优点是应用广泛，具有很高的准确性和参考意义，这在理论计算中得到了证实。为了确定其在生物传感中的潜力，提出了两种用AgBP2修饰和Ag4修饰的金纳米粒子测试Ag（I）的测试模型，其检出限分别为2 nM和75 nM。通过功能性肽（AgBP2，Ag4）的传感特性的对比，我们提供了证据，该构想可被视为敏感元件选择和性能优化的评估标准，从而在生物传感器中占据主导地位。检测限分别为2 nM和75 nM。通过功能性肽（AgBP2，Ag4）的传感特性的对比，我们提供了证据，该构想可被视为敏感元件选择和性能优化的评估标准，从而在生物传感器中占据主导地位。检测限分别为2 nM和75 nM。通过功能性肽（AgBP2，Ag4）的传感特性的对比，我们提供了证据，该构想可被视为敏感元件选择和性能优化的评估标准，从而在生物传感器中占据主导地位。